Sulfamide linkers for use in bioconjugates
a technology of sulfamide and linkers, which is applied in the field of bioconjugation, can solve the problems of limiting the choice of reactive groups, compromising conjugation efficiency, and high restrictions on biomolecules, so as to improve conjugation efficiency, reduce aggregation, and increase stability
Active Publication Date: 2020-12-29
SYNAFFIX
View PDF30 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
The invention is about a new linker that can be used to make a bioconjugate, which is a combination of two molecules. The linker can improve the efficiency of forming the bioconjugate, reduce its aggregation, increase its stability, and make it more effective as a treatment for cancer. The bioconjugate made using this linker is also a new invention.
Problems solved by technology
However, labelling of biomolecules poses high restrictions on the reaction conditions that can be applied (solvent, concentration, temperature), while the desire of chemoselective labelling limits the choice of reactive groups.
However, when no cysteine is available for conjugation, as in many proteins and certainly in other biomolecules, other methods are often required, each suffering from its own shortcomings.
However, some of the above functional groups have the disadvantage of being highly lipophilic, which may compromise conjugation efficiency, in particular in combination with a lipophilic molecule of interest (see below).
For example, a highly lipophilic linker may lead to aggregation (during and / or after conjugation), which may significantly increase reaction times and / or reduce conjugation yields, in particular when the MOI is also of hydrophobic nature.
Similarly, highly lipophilic linker-MOI combination may lead to unspecific binding to surfaces or specific hydrophobic patches on the same or other biomolecules.
However, in case of conjugation of hydrophobic, water-insoluble molecules of interest, the polarity of a PEG unit may be insufficient to offset hydrophobicity, leading to significantly reduced reaction rates, lower yields and induced aggregation issues.
In such case, lengthy PEG linkers and / or significant amounts of organic co-solvents may be required to solubilize the reagents.
In case of hydrophobic payloads, despite the use of aforementioned co-solvents, large stoichiometries of reagents may be required during conjugation while efficiency and yield may be significantly compromised due to aggregation (in process or after product isolation), as for example described by Senter et al. in Nat. Biotechn. 2014, 24, 1256-1263, incorporated by reference.
The use of long PEG spacers (12 units or more) may partially enhance solubility and / or conjugation efficiency, but it has been shown that long PEG spacers may lead to more rapid in vivo clearance, and hence negatively influence the pharmacokinetic profile of the ADC.
Using conventional linkers (e.g. PEG), effective conjugation is often hampered by the relatively low solubility of the linker-conjugate in aqueous media, especially when a relative water-insoluble or hydrophobic target molecule is used.
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
examples 1-3
of Compound 25
[0447]The synthetic approach towards compound 25 is depicted here below.
[0448]
example 1
on of Compound 21
[0449]To a solution of (R)-(+)-N-(tert-butoxycarbonyl)-O-(tert-butyldimethylsilyl)serinol (26; 50 mg, 0.164 mmol) in DCM (1.6 mL) under a nitrogen atmosphere was added chlorosulfonyl isocyanate (14.3 μL, 0.164 mmol) and the resulting mixture was stirred for 15 min. Then, Et3N (68 μL, 0.491 mmol) was added, followed by benzylamine (18 μL, 0.164 mmol). After 1 h, DCM (10 mL) and sat. NH4Cl solution (10 mL) were added. The layers were separated and the water phase was extracted with DCM (2×10 mL) and EtOAc (3×10 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. Flash chromatography (10-50% EtOAc in pentane) afforded the product (72 mg, 0.14 mmol, 85%). LRMS (ESI+) calcd for C22H39N3O7SSi (M+H+) 518.24, found 518.28.
example 2
on of Compound 22
[0450]To a solution of compound 21 (72 mg, 0.14 mmol) in DCM (1 mL) was added TFA (1 mL) and the reaction mixture was stirred at rt overnight. The reaction was concentrated and co-evaporated with toluene (2×15 mL) to afford the crude product. 1H-NMR (400 MHz, CD3OD): δ 7.38-7.24 (m, 5H), 4.29-4.18 (m, 4H), 3.91-3.64 (m, 3H), 3.54-3.43 (m, 1H), 0.98-0.86 (m, 5H), 0.15-0.04 (m, 5H). LRMS (ESI+) calcd for C17H31N3O5SSi (M+H+) 418.18, found 418.20.
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
| Property | Measurement | Unit |
|---|---|---|
| average weight distribution | aaaaa | aaaaa |
| average weight distribution | aaaaa | aaaaa |
| average weight distribution | aaaaa | aaaaa |
Login to View More
Abstract
The invention relates to a novel linker for use in bioconjugates such as antibody-drug-conjugates. The linker according to the invention is represented by formula: (I) wherein: —BM is a branching moiety; —E is a capping group; —SG is a sulfamide group; —b, c, d, e, g, i, k, l are independently 0 or 1; —f is an integer in the range of 1 to 10; —Sp1, Sp2, Sp3, Sp4, Sp5 and Sp6 are a spacer moieties; —Z1 and Z2 are connecting groups. The linker according to the invention is useful in the preparation of linker-conjugates and bioconjugates, and can be used for (a) improving conjugation efficiency in the preparation of the bioconjugate, (b) reducing aggregation during the preparation of the bioconjugate and / or of the bioconjugate, (c) increasing stability of the bioconjugate, and / or (d) increasing therapeutic index of the bioconjugate.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a U.S. national stage application of PCT / EP2017 / 052719, filed Feb. 8, 2017, which claims the benefit of and priority to European Patent Application No. 16154739.3, filed Feb. 8, 2016, the entire contents of which are incorporated herein by reference.SEQUENCE LISTING[0002]The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-WEB and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Aug. 7, 2018, is named 069818-4060 2018-08-07 _Sequence Listing.txt and is 29 KB.FIELD OF THE INVENTION[0003]The present invention is in the field of bioconjugation. The invention relates to novel sulfamide linkers and conjugates thereof, and to methods for the preparation thereof. More particularly, the invention relates to linkers comprising an acylsulfamide group and / or a carbamoyl sulfamide group and to conjugates comprising said linkers. The invention further r...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Patents(United States)
IPC IPC(8): A61K47/68C07K16/32C07K14/435C07K14/00
CPCC07K14/435A61K47/6889C07K14/00A61K47/6855A61K47/6803C07K16/32A61P35/00A61K47/68031
Inventor VAN BERKEL, SANDER SEBASTIAANHEESBEEN, RYANVERKADE, JORGE MERIJN MATHIEUWIJDEVEN, MARIA ANTONIAVAN DELFT, FLORIS LOUIS
Owner SYNAFFIX